Assay test strips with multiple labels and reading same

ABSTRACT

In one aspect, an assay test strip includes a test label that specifically binds a target analyte and a control label that is free of any specific binding affinity for the target analyte and has a different optical characteristic than the test label. In another aspect, an assay test strip includes a test label that specifically binds a target analyte and at least one non-specific-binding label that is free of any specific binding affinity for the target analyte. Systems and methods of reading assay test strips also are described.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.11/236,152, filed on Sep. 26, 2005, which is a continuation in part ofU.S. application Ser. No. 11/112,807, filed on Apr. 22, 2005, each ofwhich are incorporated by reference in their entirety.

BACKGROUND

Lateral flow assay test kits are currently available for testing for awide variety of medical and environmental conditions or compounds, suchas a hormone, a metabolite, a toxin, or a pathogen-derived antigen. FIG.1 shows a typical lateral flow test strip 10 that includes a samplereceiving zone 12, a labeling zone 14, a detection zone 15, and anabsorbent zone 20 on a common substrate 22. These zones 12-20 typicallyare made of a material (e.g., chemically-treated nitrocellulose) thatallows fluid to flow from the sample receiving zone 12 to the absorbentzone 22 by capillary action. The detection zone 15 includes a testregion 16 for detecting the presence of a target analyte in a fluidsample and a control region 18 for indicating the completion of an assaytest.

FIGS. 2A and 2B show an assay performed by an exemplary implementationof the test strip 10. A fluid sample 24 (e.g., blood, urine, or saliva)is applied to the sample receiving zone 12. In the example shown inFIGS. 2A and 2B, the fluid sample 24 includes a target analyte 26 (i.e.,a molecule or compound that can be assayed by the test strip 10).Capillary action draws the liquid sample 24 downstream into the labelingzone 14, which contains a substance 28 for indirect labeling of thetarget analyte 26. In the illustrated example, the labeling substance 28consists of an immunoglobulin 30 with a reflective particle 32 (e.g., acolloidal gold or silver particle). The immunoglobulin 30 specificallybinds the target analyte 26 to form a labeled target analyte complex. Insome other implementations, the labeling substance 28 is anon-immunoglobulin labeled compound that specifically binds the targetanalyte 26 to form a labeled target analyte complex.

The labeled target analyte complexes, along with excess quantities ofthe labeling substance, are carried along the lateral flow path into thetest region 16, which contains immobilized compounds 34 that are capableof specifically binding the target analyte 26. In the illustratedexample, the immobilized compounds 34 are immunoglobulins thatspecifically bind the labeled target analyte complexes and therebyretain the labeled target analyte complexes in the test region 16. Thepresence of the labeled analyte in the sample typically is evidenced bya visually detectable coloring of the test region 16 that appears as aresult of the accumulation of the labeling substance in the test region16.

The control region 18 typically is designed to indicate that an assayhas been performed to completion. Compounds 35 in the control region 18bind and retain the labeling substance 28. The labeling substance 28typically becomes visible in the control region 18 after a sufficientquantity of the labeling substance 28 has accumulated. When the targetanalyte 26 is not present in the sample, the test region 16 will not becolored, whereas the control region 18 will be colored to indicate thatassay has been performed. The absorbent zone 20 captures excessquantities of the fluid sample 24.

In the non-competitive-type of lateral flow assay test strip designsshown in FIGS. 2A and 2B, an increase in the concentration of theanalyte in the sample results in an increase in the concentration oflabels in the test region. Conversely, in competitive-type of lateralflow assay test strip designs, an increase in the concentration of theanalyte in the fluid sample results in a decrease in the concentrationof labels in the test region.

Although visual inspection of lateral flow assay devices of the typedescribed above are able to provide qualitative assay results, such amethod of reading these types of devices is unable to providequantitative assay measurements and therefore is prone to interpretationerrors. Automated and semi-automated lateral flow assay readers havebeen developed in an effort to overcome this deficiency.

What is needed are lateral flow assay test strips and systems andmethods of reading such test strips that provide improved detection ofdifferent capture regions on the test strips, improved assay testingspeed, and improved assay measurement sensitivity.

SUMMARY

In one aspect, the invention features an assay test strip that includesa flow path for a fluid sample and a sample receiving zone coupled tothe flow path. The assay test strip additionally includes a test labelthat specifically binds a target analyte and a control label that isfree of any specific binding affinity for the target analyte and has adifferent optical characteristic than the test label. A detection zoneis coupled to the flow path downstream of the sample receiving zone andcomprises an immobilized test reagent that specifically binds the targetanalyte and an immobilized control reagent that specifically binds thecontrol label.

In another aspect, the invention features an assay test strip thatincludes a flow path for a fluid sample, a sample receiving zone coupledto the flow path, and a detection zone coupled to the flow pathdownstream of the sample receiving zone. The assay test stripadditionally includes a test label that specifically binds a targetanalyte, and at least one non-specific-binding label that is free of anyspecific binding affinity for the target analyte. The detection zonecomprises a first immobilized reagent that specifically binds the targetanalyte and a second immobilized reagent that specifically binds thetest label, wherein each non-specific-binding label is free of anyspecific binding affinity for any of the immobilized reagents in thedetection zone.

In another aspect, the invention features a diagnostic test system thatincludes an assay test strip and a reader. The assay test strip includesa detection zone and a flow path for a fluid sample along a lateral flowdirection across the detection zone. The detection zone comprises acapture region characterized by a first dimension transverse to thelateral flow direction and a second dimension parallel to the lateralflow direction. The reader comprises an illumination system that isoperable to focus a beam of light onto an area of the detection zonehaving at least one surface dimension at most equal to smallest of thefirst and second dimensions of the capture region.

The invention also features a diagnostic test method in accordance withwhich an assay test strip is received. The lateral flow assay test stripincludes a detection zone and a flow path for a fluid sample along alateral flow direction across the detection zone. The detection zonecomprises a capture region characterized by a first dimension transverseto the lateral flow direction and a second dimension parallel to thelateral flow direction. A beam of light is focused onto one or moreareas of the detection zone having at least one surface dimension atmost equal to smallest of the first and second dimensions of the captureregion. Light intensity measurements are obtained from the illuminatedareas of the detection zone

In another aspect, the invention features a diagnostic test system thatincludes a reader and a data analyzer. The reader is operable to obtainlight intensity measurements from exposed regions of an assay test stripcomprising a detection zone. The data analyzer is operable to performoperations comprising determining a test measurement value from lightintensity measurements obtained from a test label bound to a region inthe detection zone containing a first immobilized reagent, determining acompensation measurement value from light intensity measurementsobtained from a compensation label different from the test label in oneor more regions of the detection zone free of immobilized reagents thatspecifically bind the compensation label, and determining a parametervalue from the test measurement value and the compensation measurementvalue.

The invention also features a diagnostic test method in accordance withwhich light intensity measurements are obtained from exposed regions ofan assay test strip comprising a detection zone. A test measurementvalue is determined from light intensity measurements obtained from aregion in the detection zone containing a test label bound to a firstimmobilized reagent. A compensation measurement value is determined fromlight intensity measurements obtained from a compensation labeldifferent from the test label in one or more regions of the detectionzone free of immobilized reagents that specifically bind thecompensation label. A parameter value is determined from the testmeasurement value and the compensation measurement value.

DESCRIPTION OF DRAWINGS

FIG. 1 is a diagrammatic view of a prior art implementation of a lateralflow assay test strip.

FIG. 2A is a diagrammatic view of a fluid sample being applied to asample receiving zone of the lateral flow assay test strip shown in FIG.1.

FIG. 2B is a diagrammatic view of the lateral flow assay test stripshown in FIG. 2A after the fluid sample has flowed across the test stripto an absorption zone.

FIG. 3 is a block diagram of a test strip that is loaded into anembodiment of a diagnostic test system.

FIG. 4 is a flow diagram of an embodiment of a diagnostic test method.

FIG. 5A is a diagrammatic top view of an embodiment of a test strip thatincludes separate plano-circular test and control capture regions.

FIG. 5B is a diagrammatic top view of an embodiment of a test strip thatincludes three combined test-and-control capture regions each of whichhas a plano-elliptical shape.

FIG. 6 is a diagrammatic top view of an embodiment of a test strip thatincludes a capture region that has a test reagent sub-region and acontrol reagent sub-region that are equidistant from a labeling zone.

FIG. 7A is a diagrammatic view of an implementation of the diagnostictest system shown in FIG. 3 that includes an illumination systemfocusing a light beam onto a capture region and a linear light detectorarray obtaining light intensity measurements from regions of a teststrip.

FIG. 7B is an exemplary graph of aggregate light intensity obtained bythe linear light detector array shown in FIG. 7A plotted as a functionof time.

FIG. 8 is a diagrammatic top view of an implementation of the diagnostictest system shown in FIG. 3 that includes a light source scanning alight beam over regions of a test strip.

FIG. 9 is a diagrammatic side view of an implementation of thediagnostic test system shown in FIG. 3.

FIG. 10 is a diagrammatic view of an implementation of the diagnostictest system shown in FIG. 3 that includes first and second imagingdevices that are respectively disposed directly over first and secondcapture regions of a test strip.

FIG. 11 is a diagrammatic side view of an implementation of thediagnostic test system shown in FIG. 3 obtaining measurements from thetest strip shown in FIG. 6.

FIG. 12A is a diagrammatic top view of an embodiment of a test stripthat has a labeling zone that includes a test label and anon-specific-binding label.

FIG. 12B is a diagrammatic top view of the test strip embodiment shownin FIG. 12A after a fluid sample has flowed through the test strip.

FIG. 13 is a flow diagram of an embodiment of a method executable by anembodiment of the data analyzer shown in FIG. 3.

DETAILED DESCRIPTION

In the following description, like reference numbers are used toidentify like elements. Furthermore, the drawings are intended toillustrate major features of exemplary embodiments in a diagrammaticmanner. The drawings are not intended to depict every feature of actualembodiments nor relative dimensions of the depicted elements, and arenot drawn to scale.

I . Introduction

The embodiments that are described in detail below provide lateral flowassay test strips that include multiple labels and systems and methodsfor reading such test strips that provide improved detection ofdifferent capture regions on the test strips, improved assay testingspeed, and improved assay measurement sensitivity.

The term “lateral flow assay test strip” encompasses both competitiveand non-competitive types of lateral flow assay test strips. A lateralflow assay test strip generally includes a sample receiving zone and adetection zone, and may or may not have a labeling zone. In someimplementations, a lateral flow assay test strip includes a samplereceiving zone that is located vertically above a labeling zone, andadditionally includes a detection zone that is located laterallydownstream of the labeling zone.

The term “analyte” refers to a substance that can be assayed by the teststrip. Examples of different types of analytes include organic compounds(e.g., proteins and amino acids), hormones, metabolites, antibodies,pathogen-derived antigens, drugs, toxins, and microorganisms (e.g.,bacteria and viruses).

As used herein the term “label” refers to a substance that has specificbinding affinity for an analyte and that has a detectable characteristicfeature that can be distinguished from other elements of the test strip.The label may include a combination of a labeling substance (e.g., afluorescent particle, such as a quantum dot) that provides thedetectable characteristic feature and a probe substance (e.g., animmunoglobulin) that provides the specific binding affinity for theanalyte. In some implementations, the labels have distinctive opticalproperties, such as luminescence (e.g., fluorescence) or reflectiveproperties, which allow regions of the test strip containing differentlabels to be distinguished from one another.

The term “reagent” refers to a substance that reacts chemically orbiologically with a target substance, such as a label or an analyte.

The term “capture region” refers to a region on a test strip thatincludes one or more immobilized reagents.

The term “test region” refers to a capture region containing animmobilized reagent with a specific binding affinity for an analyte.

The term “control region” refers to a capture region containing animmobilized reagent with a specific binding affinity for a label.

II. General Architecture of the Diagnostic Test System

FIG. 3 shows an embodiment of a diagnostic test system 40 that includesa housing 42, a reader 44, a data analyzer 46, and a memory 47. Thehousing 42 includes a port 48 for receiving a test strip 50. When thetest strip 50 is loaded in the port 48, the reader 44 obtains lightintensity measurements from the test strip 50. In general, the lightintensity measurements may be unfiltered or they may be filtered interms of at least one of wavelength and polarization. The data analyzer46 computes at least one parameter from one or more of the lightintensity measurements. A results indicator 52 provides an indication ofone or more of the results of an assay of the test strip 50. In someimplementations, the diagnostic test system 40 is fabricated fromrelatively inexpensive components enabling it to be used for disposableor single-use applications.

The housing 42 may be made of any one of a wide variety of materials,including plastic and metal. The housing 42 forms a protective enclosurefor the reader 44, the data analyzer 46, the power supply 54, and othercomponents of the diagnostic test system 40. The housing 42 also definesa receptacle that mechanically registers the test strip 50 with respectto the reader 44. The receptacle may be designed to receive any one of awide variety of different types of test strips 50, including test stripsof the type shown in FIG. 1.

In the illustrated embodiments, each of the test strips 50 is anon-competitive type of lateral flow assay test strip that supportslateral flow of a fluid sample along a lateral flow direction 51 andincludes a labeling zone containing a labeling substance that binds alabel to a target analyte and a detection zone that includes at leastone test region containing an immobilized substance that binds thetarget analyte. One or more areas of the detection zone, including atleast a portion of the test region, are exposed for optical inspectionby the reader 44. The exposed areas of the detection zone may or may notbe covered by an optically transparent window.

In other embodiments, the test strips are competitive type of lateralflow assay test strips in which the concentrations of the label in thetest region decreases with increasing concentration of the targetanalyte in the fluid sample. Some of these embodiments include alabeling zone, whereas others of these implementations do not include alabeling zone.

Some of these competitive lateral flow assay test strip embodimentsinclude a labeling zone that contains a label that specifically bindstarget analytes in the fluid sample, and a test region that containsimmobilized target analytes as opposed to immobilized test reagents(e.g., antibodies) that specifically bind any non-bound labels in thefluid sample. In operation, the test region will be labeled when thereis no analyte present in the fluid sample. However, if target analytesare present in the fluid sample, the fluid sample analytes saturate thelabel's binding sites in the labeling zone, well before the label flowsto the test region. Consequently, when the label flows through the testregion, there are no binding sites remaining on the label, so the labelpasses by and the test region remains unlabeled.

In other competitive lateral flow assay test strip embodiments, thelabeling zone contains only pre-labeled analytes (e.g., gold adhered toanalyte) and the test region contains immobilized test reagents with anaffinity for the analyte. In these embodiments, if the fluid samplecontains unlabeled analyte in a concentration that is large compared tothe concentration of the pre-labeled analyte in the labeling zone, thenlabel concentration in the test region will appear proportionatelyreduced.

The reader 44 includes one or more optoelectronic components foroptically inspecting the exposed areas of the detection zone of the teststrip 50. In some implementations, the reader 44 includes at least onelight source and at least one light detector. In some implementations,the light source may include a semiconductor light-emitting diode andthe light detector may include a semiconductor photodiode. Depending onthe nature of the label that is used by the test strip 50, the lightsource may be designed to emit light within a particular wavelengthrange or light with a particular polarization. For example, if the labelis a fluorescent label, such as a quantum dot, the light source would bedesigned to illuminate the exposed areas of the detection zone of thetest strip 50 with light in a wavelength range that induces fluorescentemission from the label. Similarly, the light detector may be designedto selectively capture light from the exposed areas of the detectionzone. For example, if the label is a fluorescent label, the lightdetector would be designed to selectively capture light within thewavelength range of the fluorescent light emitted by the label or withlight of a particular polarization. On the other hand, if the label is areflective-type label, the light detector would be designed toselectively capture light within the wavelength range of the lightemitted by the light source. To these ends, the light detector mayinclude one or more optical filters that define the wavelength ranges orpolarizations axes of the captured light.

The data analyzer 46 processes the light intensity measurements that areobtained by the reader 44. In general, the data analyzer 46 may beimplemented in any computing or processing environment, including indigital electronic circuitry or in computer hardware, firmware, orsoftware. In some embodiments, the data analyzer 46 includes a processor(e.g., a microcontroller, a microprocessor, or ASIC) and ananalog-to-digital converter. In the illustrated embodiment, the dataanalyzer 46 is incorporated within the housing 42 of the diagnostic testsystem 40. In other embodiments, the data analyzer 46 is located in aseparate device, such as a computer, that may communicate with thediagnostic test system 40 over a wired or wireless connection.

In general, the results indicator 52 may include any one of a widevariety of different mechanisms for indicating one or more results of anassay test. In some implementations, the results indicator 52 includesone or more lights (e.g., light-emitting diodes) that are activated toindicate, for example, a positive test result and the completion of theassay test (i.e., when sufficient quantity of labeling substance 28 hasaccumulated in the control region). In other implementations, theresults indicator 52 includes an alphanumeric display (e.g., a two orthree character light-emitting diode array) for presenting assay testresults.

A power supply 54 supplies power to the active components of thediagnostic test system 40, including the reader 44, the data analyzer46, and the results indicator 52. The power supply 54 may be implementedby, for example, a replaceable battery or a rechargeable battery. Inother embodiments, the diagnostic test system may be powered by anexternal host device (e.g., a computer connected by a USB cable).

FIG. 4 shows an embodiment of a diagnostic test method that isexecutable by implementations of the diagnostic test system 40 describedbelow. In accordance with this method, the reader 44 obtains separablelocalized light intensity measurements from regions of the exposed areaof the detection zone of the test strip 50 when the test strip 50 isloaded in the port 48 of the diagnostic test system 40 (block 60.

As used herein, the term “separable localized light intensitymeasurements” refers to the ability of the reader 44 transmit or recordthe light intensity measurements from respective localized regions ofthe test strip in a way that allows the data analyzer 46 to individuallyanalyze each of the light intensity measurements.

In this embodiment, each of the separable localized regions from whichthe light intensity measurements are obtained by the reader 44 ischaracterized by at least one surface dimension that is smaller than thedimension of the exposed area of the detection zone that is transverseto the lateral flow direction. In some implementations, each of theselocalized regions has a surface dimension that is approximately the samesize or smaller than the narrowest dimension of a region of interest inthe detection zone (e.g., the test region, the control region, or aregion of an immobilized labeled or unlabeled complex).

After the reader 44 has obtained light intensity measurements from suchlocalized regions of interest in the detection zone (block 60), the dataanalyzer 46 identifies ones of the light intensity measurements obtainedfrom the regions of interest (block 62). In this process, the dataanalyzer 46 isolates the measurements corresponding to regions ofinterest from the measurements corresponding to other regions of thetest strip 50. The isolated measurements have higher signal-to-noiseratios than aggregated measurements that include measurements fromregions outside of the regions of interest.

The data analyzer 46 computes at least one parameter from ones of theidentified light intensity measurements (block 64). Exemplary parametersinclude peak intensity and aggregate intensity values. Since themeasurements that are used to compute these parameters have highersignal-to-noise ratios, they characterize the region of interest withgreater accuracy and, thereby, improve the results of the lateral flowassay.

III. Test Strips with Different Test and Control Labels and Reading theSame

The embodiments that are described in this section provide lateral flowassay test strips that include different labels for test and control, aswell as systems and methods of reading such test strips. The use ofdifferent test and control labels enables the test strip and thediagnostic test system to be designed in ways that improve thedetectability of the test and control signals, improve assay testingspeed, and improve assay measurement sensitivity.

A. Test Strips Having Different Test and Control Labels

In these embodiments, the labeling zone 14 includes a test label and acontrol label for each analyte to be assayed. In general, the test labeland the associated control label have different respective detectablecharacteristics or properties. In some implementations, the test labelfluoresces at a first characteristic wavelength and the control labelfluoresces at a second characteristic wavelength different from thefirst characteristic wavelength. The test label may include, forexample, quantum dots of a first type and the control label may include,for example, quantum dots of a second type. In some implementations, thequantum dots are nanometer sized semiconductor nanocrystals withfluorescent properties that are determined by their sizes. In this way,the fluorescence wavelengths of the quantum dots can be tuned bychanging the sizes of the quantum dots.

The test label is bound by an immobilized test reagent in the detectionzone 15 that has a specific binding affinity for the test label. Thecontrol label is bound by an immobilized control reagent in thedetection zone 15 that has a specific binding affinity for the controllabel. The different test and control labels may be immobilized in thedetection zone 15 of the test strip in separate discrete capture regionsor in the same test-and-control capture region. In general, the test andcontrol regions may have any shape, including rectangular andnon-rectangular shapes.

For example, FIG. 5A shows an embodiment of the test strip 50 that has atest region 70 that contains an immobilized test reagent and a separateand distinct control region 72 that contains an immobilized controlreagent. In the illustrated embodiment, each of the test and controlregions 70, 72 has a respective plano-circular shape in the plane of thesurface of the detection zone 15. Such a shape allows a suitablyconfigured reader 44 to project an illumination beam onto aplano-circular area of the detection zone 15 that substantiallycorresponds to the test-and-control regions 70, 72 in size and shape. Inthis way, the light intensity measurements that are obtained from thetest and control labels may have higher signal-to-noise levels,increased measurement sensitivity, and reduced incidence of erroneousresults for low concentrations of analytes.

FIG. 5B shows an exemplary implementation of the test strip 50 thatincludes three combined test-and-control regions 74, 76, 78. Each of thetest-and-control regions 74-78 includes a respective immobilized testreagent and a respective immobilized control reagent that are intermixedin the same region of the detection zone 15. Each test reagent has aspecific binding affinity for a respective target analyte that isspecifically bound by a respective test label in the labeling zone 14and each control reagent has a specific binding affinity for arespective control label in the labeling zone 14. In someimplementations, the test label and the control label in each captureregion 74-78 fluoresce at different characteristic wavelengths, enablingtheir fluorescent emissions to be distinguished using wavelengthseparation techniques (e.g., diffractive optics and optical filters).

In the test strip embodiment shown in FIG. 5B, each test-and-controlregion 74-78 has a respective plano-elliptical shape in the plane of thesurface of the detection zone 15. Such a shape allows a suitablyconfigured reader 44 to project an illumination beam onto aplano-elliptical area of the detection zone 15 that substantiallycorresponds to the test-and-control regions 74-78 in size and shape. Inthis way, the light intensity measurements that are obtained from thetest and control labels may have higher signal-to-noise levels,increased measurement sensitivity, and reduced the incidence oferroneous results for low concentrations of analytes.

FIG. 6 shows an implementation of the test strip 50 in which the testreagent and the control reagent are immobilized in respectivesub-regions 80, 82 of a combined test-and-control region 84. In theillustrated embodiment, the test reagent sub-region 80 and the controlreagent sub-region 82 are equidistant from the labeling zone 14 (i.e.,D_(TEST)=D_(CONTROL)). The test reagent sub-region 80 and the controlreagent sub-region 82 also are equidistant from the sample receivingzone 12. In this implementation, the fluid sample need only flow throughone capture location along the lateral flow direction before the testand control results can be determined. Therefore, the assay testingspeed with respect to this test strip implementation may be faster thanthe assay testing speed for implementations in which separate test andcontrol regions are located at different locations in the lateral flowdirection 51.

B. Illuminating the Different Test and Control Labels

The illumination source of the reader 44 may be constructed and arrangedto focus or scan a light beam onto an area of the detection zone that issized and shaped so that the light reflected or fluorescing from thetest and control labels easily may be segmented from other regions ofthe detection zone 15. In some embodiments, the capture regions (i.e.,the separate test and control regions and the combined test-and-controlregions) may have non-rectangular shapes, which allow relativelyinexpensive optical components to be used to project light beams ontoareas of the detection zone 15 that substantially correspond to thecapture regions in size and shape. In this way, the aggregate signalsthat are generated by the detection system of the reader 44 largelycorresponds to light respectively reflecting or fluorescing from theilluminated capture regions and therefore have higher signal-to-noiseratios than comparable measurements obtained as a result of illuminationof areas that do not substantially correspond to the shapes of thecapture regions.

FIG. 7A shows an implementation of the test strip 50 and animplementation of the diagnostic test system 40.

The test strip includes a test region 90 that is separate and distinctfrom the control region 92. The test region 90 includes an immobilizedtest reagent that specifically binds a target analyte and the controlregion 92 includes an immobilized control reagent that specificallybinds a label. Each of the test and control capture regions 90, 92 ischaracterized by a first dimension (W_(T), W_(C)) transverse to thelateral flow direction 51 and a second dimension (L_(T), L_(C)) parallelto the lateral flow direction 51.

The diagnostic test system 40 includes a light source 96, a linear array98 of light detectors 100, and a lens 102. In this implementation, thediagnostic test system 40 additionally includes a mechanism (not shown)for imparting relative motion between the optical inspection components104 (e.g., the light source 96, the lens 102, and the linear lightdetector array 98) and the test strip 50. The motion-imparting mechanismmay be any one of a wide variety of different mechanisms, including amotorized carriage that moves the optical inspection components relativeto the test strip 50 on a pair of rails, and one or more motorized drivewheels that move the test strip 50 relative to the optical inspectioncomponents 104. In the illustrated embodiment, the optical inspectioncomponents 104 are shown moving relative to the test strip 50 in thedirection of arrow 105 (i.e., in the lateral flow direction 51).

The linear light detector array 98 is oriented in a direction transverseto the direction of motion of the optical inspection components 104.

In implementations in which the test and control labels aredistinguishable by, for example, their different wavelengths offluorescent emission, the test and control regions 90, 92 may coincide.In these implementations, multiple optical filters or a single tunableoptical filter may be used to distinguish the light that is receivedfrom the test and control labels.

In operation, the light source 96 illuminates an area of the exposedportion of the detection zone 15 with a beam of light 106 as the opticalinspection components 104 are moved relative to the test strip 50. Theilluminating light may be broadband or narrowband and may be polarizedor non-polarized. The light source 96 focuses the light beam 106 ontothe detection zone 15 with a shape having at least one surface dimensionthat is at most the smaller of characteristic dimensions (W_(T), L_(T),W_(C), L_(C)) of the test and control capture regions 90, 92. In theillustrated embodiment, the light beam 106 illuminates an area thatsubstantially corresponds to each of the capture regions in size andshape.

The linear light detector array 98 obtains separable localized lightintensity measurements from a narrow portion of the illuminated regionof the detection zone 15. In general, the light intensity measurementsmay be unfiltered or they may be filtered in terms of wavelength orpolarization. The light detector array 98 may be synchronized with thelight source 96. In general, the light detector array 98 may measurelight intensity while the detection zone 15 is being illuminated orafter the light source 96 has illuminated the detection zone 15. Lightreflected or fluorescing from the detection zone 15 is focused by thelens 102 onto the individual light detectors 100 of the light detectorarray 98. Each of the light detectors 100 receives light from arespective localized region of the detection zone 15. That is, eachlight detector 100 is able to resolve or separably image a respectivelocalized region of the detection zone 15. The light detectors 100produce signals representative of the amount of light received from therespective localized regions. These signals may be stored in a memory orthey may be transmitted to the data analyzer 46 for processing.

When the illuminated area of the detection zone 15 coincides with one ofthe capture regions 90, 92, the aggregate signal that is generated bythe light detector array 98 largely corresponds to light reflecting orfluorescing from the illuminated capture region and therefore has ahigher signal-to-noise ratio than comparable measurements obtained fromlarger areas of the detection zone 15. The test strip 50 may includeposition markers or other features that may be used by the diagnostictest system 40 to determine when the illuminated area of the zone 15coincides with one of the capture regions 90, 92. A description ofexemplary types of alignment features may be obtained from U.S. patentapplication Ser. No. 11/280,640, filed Nov. 16, 2005, by Patrick T.Petruno et al. and entitled “ASSAY TEST STRIPS AND READING SAME”.

The data analyzer 46 (FIG. 3) is operable to process the signals thatare generated by the individual light detectors 100 of the linear array98 to identify the ones of the light intensity measurements that areobtained from the regions of interest (e.g., the test region 90 and thecontrol region 92). In some implementations, the surface of thedetection zone 15 is substantially homogeneous in the directiontransverse to the lateral flow direction. In these implementations, thesignals from the light detectors in the linear array 98 may beaggregated without substantial loss of information.

FIG. 7B shows an exemplary graph 108 of the aggregated intensitymeasurements that are produced by the linear light detector array 98plotted as a function of time. In this example, the graph 108 includeshigher intensity aggregate intensities 110, 112 when the light detectors100 in the array 98 are positioned over the test region 90 and thecontrol region 92. With respect to this example, the data analyzer 46may identify the light intensity measurements that are obtained from thetest region 90 and the control region 92 by thresholding the graph 108at an intensity threshold level 114. The light intensity measurementsthat are above the threshold level 114 are identified as having comefrom the test region 90 and the control region 92. Additionalinformation, such as the relative times the identified ones of the lightintensity measurements were obtained, may be used by the data analyzer46 to correlate the identified light intensity measurements with thetest region 90 and the control region 92.

FIG. 8 shows an implementation of the diagnostic test system 40 thatincludes a light source 120 that is operable to scan a light beam 122across the exposed area of the detection zone 15. The light source 120focuses the light beam 122 onto the detection zone 15 with a shapehaving at least one surface dimension that is at most the smaller ofcharacteristic dimensions (W_(T), L_(T), W_(C), L_(C)) of the test andcontrol capture regions 90, 92. In the illustrated embodiment, thesurface area illuminated by the light beam 122 has a circular dimensionthat is smaller than each of the characteristic dimensions (W_(T),L_(T), W_(C), L_(C)) of the test and control regions 90, 92. The lightbeam 122 may be broadband or narrowband and may be polarized ornon-polarized.

In general, the light source 120 may scan the light beam 122 across theexposed area of the detection zone 15 along any path that includes thetest region 90 and the control region 92, including in a directiontransverse to the lateral flow direction and a direction parallel to thelateral flow direction. In the illustrated embodiment, the light source120 scans the light beam 122 along a circuitous zigzag path 124 acrossthe exposed area of the detection zone 15. In some implementations, thelight source 120 includes a light emitter, such as a light-emittingdiode or a laser, and one or more optical components (e.g., one or morelenses and a rotating mirror) for shaping and scanning the emitted lightto produce the beam 122.

In the implementation shown in FIG. 8, the diagnostic test system 40 mayobtain separable localized light intensity measurements using any typeof single-element or multi-element light detector that has a field ofview that encompasses the path of the light beam 122 across the exposedarea of the detection zone 15 or that tracks the localized regions ofthe detection zone 15 as they are illuminated by the light beam 122. Thelight intensity measurements may be unfiltered or they may be filteredin terms of wavelength or polarization. The light detector array may besynchronized with the light source. In general, the light detector arraymay measure light intensity while individual separable localized regionsof the detection zone 15 are being illuminated or after the light sourcehas illuminated the individual separable localized regions of thedetection zone 15. Because the light beam 122 illuminates only a singlelocalized region of the detection zone 15 at a time, the light obtainedby the light detector corresponds to the light reflected or fluorescingfrom the illuminated localized region. Therefore, s each data point ofthe signal that is generated by the light detector correlates with arespective localized region and has a higher signal-to-noise ratio thancomparable measurements obtained from larger regions of the detectionzone 15.

The data analyzer 46 (FIG. 3) is operable to process the signals thatare generated by the light detectors to identify the ones of the lightintensity measurements that are obtained from the regions of interest(e.g., the test region 90 and the control region 92). For example, insome implementations, the data analyzer 46 may identify the lightintensity measurements that are obtained from the test region 90 and thecontrol region 92 by thresholding the time-varying light intensitymeasurement signal that is generated by the light detector. The ones ofthe light intensity measurements that are above the threshold level areidentified as having come from the test region 90 and the control region92. Additional information, such as the relative times the identifiedones of the light intensity measurements were obtained, may be used bythe data analyzer 46 to correlate the identified light intensitymeasurements with the test region 90 and the control region 92.

In the implementation of the test strip 50 that is shown in FIG. 8, aset of regularly spaced position markers 126 are located along one edgeof the test strip 50. The position markers 126 include features thathave a different optical characteristic (e.g., reflectivity orfluorescence) than the surface of the test strip 50. As a result, themeasurements obtained near the edge of the test strip 50 vary inintensity in accordance with the pattern of the position markers 126. Inthis way, the position markers encode positions along the test strip 50in the lateral flow direction. The data analyzer 46 may determine theencoded positions along the lateral flow direction by incrementing aposition counter with each intensity variation cycle (e.g.,peak-to-valley) in the light intensity measurements obtained from theedge of the detection zone 15. In these implementations, the dataanalyzer 46 correlates the light intensity measurements with thepositions along the test strip 50 in the lateral flow direction 51. Thelocation correlation information may be stored in a lookup table that isindexed by the position counter value. Based on this information andpredetermined information correlating the locations of the regions ofinterest with the light intensity contrast pattern produced by theposition markers 126, the data analyzer 46 can identify the ones of thelight intensity measurements corresponding to the regions of interest.

C. Detecting Light from the Different Test and Control Labels

In some implementations, the test label fluoresces at a firstcharacteristic wavelength and the control label fluoresces at a secondcharacteristic wavelength different from the first characteristicwavelength. Implementations of the reader 44 may include a detectionsystem that is constructed and arranged to segment the light reflectedor fluorescing from the test and control labels from each other and fromother regions of the detection zone 15.

FIG. 9 shows an implementation of the reader 44 that may be used toseparate the fluorescent light emitted from the test and control labelsthat are bound in separate test and control regions (see, e.g., FIG. 5A)or in the same test-and-control region (see, e.g., FIGS. 5B and 6). Thereader 44 obtains separable localized light intensity measurements usinga light detector array 160, wherein each individual detector element162, 164 is the target of a specific characteristic wavelength of lightthat is separated and steered by a diffractive lens 166. In theillustrated example, the different test and control labels areimmobilized in a combined test-and-control region 168. The light 170from the immobilized test label is steered by the diffractive lens 166to the detector element 162, whereas the light from the immobilizedcontrol label is steered by the diffractive lens to the detector element164. In the implementation shown in FIG. 9, each detector element 162,164 additionally may include a respective optical filter with narrowpassbands selected to preferentially transmit light from the test andcontrol labels, respectively.

In some embodiments, the reader 44 includes first and second opticalfilters that are constructed and arranged to filter light received byfirst and second light detectors (or first and second regions of ashared light detector array). The first optical filter selectivelytransmits light within a first wavelength range that encompasses thefirst characteristic wavelength and excludes the second characteristicwavelength. The second optical filter selectively transmits light withina second wavelength range that encompasses the second characteristicwavelength and excludes the first characteristic wavelength. In someimplementations, the first and second wavelength ranges substantiallycorrespond to the predominant fluorescent emission spectra of the testand control labels. In this way, the first and second optical filtersare able to reduce noise that might be caused by light from elementsother than the test and control labels, respectively. In someimplementations, the optical filters are polarized to selectivelytransmit polarized light that is received from the test and controllabels.

With respect to test strip implementations in which the different testand control labels are immobilized in separate discrete test and controlregions in the detection zone 15, the test and control labels may beassayed by embodiments of the diagnostic test system 40 in which thepositions of the detector components are correlated with the positionsof the test and control regions to improve the ability to detect thetest and control regions of the test strip.

For example, FIG. 10 shows an embodiment of the test strip 50 thatincludes a test region 90 and a control region 92, which respectivelycontain immobilized test and control reagents. The test and controlreagents respectively have specific binding affinity for test andcontrol labels in the labeling zone 14. In some implementations, thetest label fluoresces at a first characteristic wavelength and thecontrol label fluoresces at a second characteristic wavelength differentfrom the first characteristic wavelength. The test label may include,for example, quantum dots of a first type and the control label mayinclude, for example, quantum dots of a second type.

FIG. 10 also shows an implementation of the diagnostic test system 40that includes a light source 180 and a pair of light detectors 182, 184.The light source 180 may be implemented by one or more light-emittingdiodes that generate a relatively broad beam of light that illuminatesthe regions of interest in the detection zone 15. The light detectors182, 184 may be implemented by single-element light detectors ormulti-element light detectors that are disposed directly over the testregion 90 and the control region 92 when the test strip 50 is loadedwithin the port 48 of the diagnostic test system 40. The light detectors182, 184 respectively include first and second optical filters 186, 188.The first optical filter 186 selectively transmits light within a firstwavelength range that encompasses the first characteristic wavelengthand excludes the second characteristic wavelength. The second opticalfilter 188 selectively transmits light within a second wavelength rangethat encompasses the second characteristic wavelength and excludes thefirst characteristic wavelength. In some implementations, the first andsecond wavelength ranges substantially correspond to the predominantfluorescent emission spectra of the test and control labels. In thisway, the optical filters 186, 188 are able to reduce noise that might becaused by light from elements other than the test and control labels,respectively. In some implementations, the optical filters 186, 188 arepolarized to selectively transmit polarized light received from the testand control labels.

In operation, the light source 180 illuminates the test region 90 andthe control region 92 with light 190. The illuminating light 190 may bebroadband or narrowband and may be polarized or non-polarized. The lightdetectors 182, 184 obtain separable localized light intensitymeasurements from the illuminated regions of the detection zone 15. Thedetectors 182, 184 may be synchronized with the light source 180. Ingeneral, the light detectors 182, 184 may measure light intensity whilethe detection zone 15 is being illuminated or after the light source 180has illuminated the detection zone 15. Light reflected or fluorescingfrom the test region 90 and the control region 92 is focused by thelenses 187, 189 onto the light detectors 182, 184, respectively. In thisway, the light detectors 182, 184 are able to resolve or separably imagethe test region 90 and the control region 92. The light detectors 182,184 produce signals representative of the amount of light received fromthe test region 90 and the control region 92. If the light detectors182, 184 are implemented by single-element detectors, the signalsrepresent total or aggregate amounts of light received from the testregion 90 or the control region 92. If the light detectors 182, 184 areimplemented by multi-element detectors, the signals represent theamounts of light received from localized areas of the test region 90 andthe control region 92. The signals that are generated by the lightdetectors 182, 184 may be stored in a memory or they may be transmittedto the data analyzer 46 for processing.

In some implementations, the light reflected or fluorescing from thetest region 90 and the control region 92 is preferentially transmittedthrough apertures 192, 194 in an aperture plate 196, whereas light fromother regions of the test strip 50 are substantially blocked by theaperture plate 196. As a result, the signals that are generated by thelight detectors 182, 184 have higher signal-to-noise ratios thancomparable measurements obtained from larger regions of the detectionzone 15. In addition, the light obtained by the light detectors 182, 184substantially corresponds to the light reflected or fluorescing from thetest region 90 and the control region 92, respectively. Therefore, thesignals that are generated by the light detectors 182, 184 correlatewith the test region 90 and the control region 92, respectively, and thedata analyzer 46 can identify the ones of the light intensitymeasurements that are obtained from the test region 90 and the controlregion 92 directly. That is, the light intensity measurements generatedby the light detector 182 are obtained from the test region 90 and thelight intensity measurements generated by the light detector 184 areobtained from the control region 92.

FIG. 11 shows an implementation of the diagnostic test system 40 thatincludes an illumination system 200, which has a first detector 202, asecond detector 204, and a light source 206.

The light source 206 may be implemented by one or more light-emittingdiodes that generate a relatively broad beam of light 207 thatilluminates the regions of interest in the detection zone 15. The lightdetectors 202, 204 may be implemented by single-element light detectorsor multi-element light detectors that are disposed directly over thetest reagent sub-region 80 and the control reagent sub-region 82 whenthe test strip 50 is loaded within the port 48 of the diagnostic testsystem 40. The light detectors 202, 204 respectively include first andsecond optical filters 208, 210. The first optical filter 208selectively transmits light within a first wavelength range thatencompasses the first characteristic wavelength and excludes the secondcharacteristic wavelength. The second optical filter 210 selectivelytransmits light within a second wavelength range that encompasses thesecond characteristic wavelength and excludes the first characteristicwavelength. In some implementations, the first and second wavelengthranges substantially correspond to the predominant fluorescent emissionspectra of the test and control labels. In this way, the optical filters208, 210 are able to reduce noise that might be caused by light fromelements other than the test and control labels, respectively. In someimplementations, the optical filters 208, 210 are polarized toselectively transmit polarized light received from the test and controllabels.

In operation, the light source 206 illuminates the test reagentsub-region 80 and the control reagent sub-region 82 with light 212. Theilluminating light 207 may be broadband or narrowband and may bepolarized or non-polarized. The light detectors 202, 204 obtainseparable localized light intensity measurements from the illuminatedregions of the detection zone 15. The detectors 202, 204 may besynchronized with the light source 206. In general, the light detectors202, 204 may measure light intensity while the detection zone 15 isbeing illuminated or after the light source 200 has illuminated thedetection zone 15. Light reflected or fluorescing from the test reagentsub-region 80 and the control reagent sub-region 82 is focused by thelenses 214, 216 onto the light detectors 202, 204, respectively. In thisway, the light detectors 202, 204 are able to resolve or separably imagethe test reagent sub-region 80 and the control reagent sub-region 82.The light detectors 202, 204 produce signals representative of theamount of light received from the test reagent sub-region 80 and thecontrol reagent sub-region 82. If the light detectors 202, 204 areimplemented by single-element detectors, the signals represent total oraggregate amounts of light received from the test reagent sub-region 80or the control reagent sub-region 82. If the light detectors 202, 204are implemented by multi-element detectors, the signals represent theamounts of light received from localized areas of the test reagentsub-region 80 and the control reagent sub-region 82. The signals thatare generated by the light detectors 202, 204 may be stored in a memoryor they may be transmitted to the data analyzer 46 for processing.

IV. Test Strips with Non-Specific-Binding Labels and Reading the Same

The embodiments that are described in this section provide lateral flowassay test strips that include one or more compensation labels that arenot specifically bound by the regions from which compensationmeasurements are made. A compensation label may bind non-specifically tothe capture and non-capture regions of the detection zone 15, or it maybind specifically to some but not all of the regions of the detectionzone 15. These embodiments include implementations of the diagnostictest system 40 that are designed to compensate for non-specific bindingeffects in parameter values that are derived from light intensitymeasurements of test and/or control labels based on light intensitymeasurements obtained from the compensation labels. These embodimentsmay be implemented with respect to both competitive and non-competitivetypes of lateral flow assay test strips embodiments.

FIGS. 12A and 12B show an implementation of the test strip 50 in whichthe labeling zone 14 includes a test label 230, a control label 232, anda non-specific-binding compensation label 234. The detection zone 15includes a test region 90 that includes an immobilized test reagent thatspecifically binds a target analyte, and a control region 92 thatincludes an immobilized control reagent that specifically binds thecontrol label. In some implementations, each of the labels 230-234fluoresces at a different characteristic wavelength. The labels 230-234may include, for example, quantum dots of different respective types.

FIG. 12A shows the test strip 50 before a fluid sample has been appliedto the sample receiving zone 12. FIG. 12B shows the test strip 50 aftera fluid sample, which was applied to the sample receiving zone 12, hasflowed along the flow path in the lateral flow direction 51 from thesample receiving zone 12 to the absorbent zone 20. In the illustratedexample, the applied fluid sample contained the target analyte that isspecifically bound by the test label and the test reagent.

As shown in FIG. 12B, the test label 230 accumulates predominantly inthe test region 90 and the control label 232 accumulates predominantlyin the control region 92, although both the test label 230 and thecontrol label 232 exhibit non-specific binding in the non-captureregions of the detection zone 15. The non-specific-binding compensationlabel 234, on the other hand, is distributed uniformly across thedetection zone 15 due to the non-specific binding characteristics of thenon-specific-binding compensation label.

In general, any of the implementations of the diagnostic test systemthat are described above may be used to obtain light intensitymeasurements from the various regions of the implementation of the teststrip 50 that is shown in FIG. 12B. In some implementations, the dataanalyzer 46 (FIG. 3) compensates for the effects of non-specific bindingon the measurements of the test and control labels 230, 232 in the testand control regions 90, 92 based on light intensity measurementsobtained from the non-specific-binding compensation label 234.

FIG. 13 shows a flow diagram of an embodiment of a method by which thedata analyzer 46 estimates the effects of non-specific binding on lightintensity measurements that are obtained from a test label.

In accordance with this embodiment, the data analyzer 46 (FIG. 3)determines a test measurement value from light intensity measurementsobtained from a test region containing an immobilized test reagent thatspecifically binds the target analyte (block 240). In someimplementations, the test measurement value corresponds to a statisticalmeasure (e.g., a peak intensity value or average intensity value) thatis computed from the light intensity measurements that are obtained fromthe test region.

The data analyzer 46 also determines a compensation measurement valuefrom light intensity measurements obtained from a compensation label inone or more regions of the detection zone 15 that are free ofimmobilized reagents that specifically bind the compensation label(block 242).

In general, the compensation label may be any label that is notspecifically bound in the region from which the compensation measurementvalue is determined. For example, in some implementations, thecompensation label may be the non-specific binding label 234. In otherimplementations, the compensation label may be the control label 232when the compensation measurement is determined based on light intensitymeasurements obtained from a region (e.g., the test region) that is freeof immobilized reagents that specifically bind the control label. Inimplementations in which the compensation label has a characteristic orproperty that is distinguishable from the test label and/or the controllabel (e.g., by different wavelengths of fluorescent emission), thecompensation measurement may be determined from light intensitymeasurements obtained from the test and/or control regions. With respectto these implementations, the data analyzer 46 may determine respectivecompensation measurements from light intensity measurements obtainedfrom within the regions of interest (e.g., the test and control regions)and from regions outside of the regions of interest. The data analyzer46 may use these different compensation measurements in thedetermination of the evaluation parameter value.

The compensation measurement value may correspond to a statisticalmeasure (e.g., a peak intensity value or average intensity value) thatis computed from one or more regions of the detection zone that are freeof immobilized reagents. In some implementations, the compensationmeasurement value is determined from light intensity measurements thatare obtained from regions that are immediately adjacent to the testregion. In other implementations, the compensation measurement valuecorresponds to an average light intensity value that is determined fromlight intensity measurements that are obtained from non-capture regionsacross the detection zone 15.

The data analyzer 46 (FIG. 3) determines a parameter value from the testmeasurement value and the compensation measurement value (block 244).The parameter value may be, for example, a relative quantity of thetarget analyte or an absolute quantity of the target analyte. A relativequantity of the target analyte may be determined by comparing (e.g., bydifference or ratio operations) the test measurement value to thecompensation measurement value.

An absolute quantity of the target analyte may be determined bycorrecting the test measurement value based on the compensationmeasurement value and comparing the corrected test measurement value toa calibration table that maps corrected test measurement values totarget analyte quantities. The calibration table may be derived by thedata analyzer 46 from light intensity measurements obtained from one ormore calibration regions that may be included on the test strip.

In some implementations, the data analyzer 46 (FIG. 3) may determine anevaluation parameter value based on the determined test measurementvalue and the determined compensation measurement value. The evaluationparameter may correspond, for example, to a value (e.g., yes/no, orpositive/negative) that indicates the result of an assay test. In someimplementations, the data analyzer 46 compares the ratio of the testmeasurement value (M_(T)) and the compensation measurement value (M_(C))to a threshold value (T1) to determine the evaluation parameter value(EP1). For example,If (M _(T))/(M _(C))>T1, EP1=1,otherwise, EP1=0.

In another implementation, the data analyzer 46 compares the differencebetween the test measurement value (M_(T)) and the compensationmeasurement value (M_(C)) to a threshold value (T2) to determine theevaluation parameter value (EP2). For example,If (M _(T))−(M _(C))>T2, EP2 =1,otherwise, EP2=0.

In some embodiments, the data analyzer 46 (FIG. 3) may determinerelative quantities of two different analytes (A, B) in a fluid samplebased on test measurement values that are determined from lightintensity measurements obtained from respective test regions withimmobilized test reagents with respective binding affinities for thetarget analytes A, B. In these embodiments, the relative quantities(MR_(A), MR_(B)) may correspond to test measurement values (M_(A),M_(B)) that have been corrected by a determined compensation measurementis value (M_(C)). For example,MR _(A) =M _(A) −M _(C)MR _(B) =M _(B) −M _(C)

In some implementations, the data analyzer 46 (FIG. 3) compares theratio of the corrected test measurement values (M_(A)−M_(C),M_(B)−M_(C)) to a threshold value (T3) to determine an evaluationparameter value (EP3). For example,If (M _(A) −M _(C))/(M _(B) −M _(C))>T3, EP3=1,otherwise, EP3=0.V. Conclusion

The embodiments that are described above provide lateral flow assay teststrips that include multiple labels and systems and methods for readingsuch test strips that provide improved detection of different captureregions on the test strips, improved assay testing speed, and improvedassay measurement sensitivity.

Other embodiments are within the scope of the claims.

What is claimed is:
 1. A diagnostic test system, comprising: an assaytest strip comprising a flow path for a fluid sample; a sample receivingzone coupled to the flow path; a test label that specifically binds atarget analyte; a control label that is free of any specific bindingaffinity for the target analyte and has a different opticalcharacteristic than the test label; and a detection zone coupled to theflow path downstream of the sample receiving zone, the detection zonecomprising a capture region; an immobilized test reagent thatspecifically binds the target analyte in the capture region; and animmobilized control reagent that specifically binds the control label inthe capture region, wherein the immobilized test reagent and theimmobilized control reagent are intermixed in the capture region; and areader comprising an illumination system that focuses a beam of lightonto an area of the detection zone, the cross-section of the focusedbeam of light on the surface of the detection zone having a size andshape substantially corresponding to the size and shape of the captureregion.
 2. The system of claim 1, wherein the capture region has aplano-elliptical shape, and the illumination system is operable to focusa beam of light onto the plano-elliptical area of the capture region. 3.The system of claim 1, wherein the test label fluoresces at a firstcharacteristic wavelength and the control label fluoresces at a secondcharacteristic wavelength different from the first characteristicwavelength.
 4. The system of claim 1, wherein the test label comprisesquantum dots of a first type and the control label comprises quantumdots of a second type.
 5. The system of claim 1, wherein the assay teststrip comprises a set of regularly spaced position markers along an edgeof the assay test strip, and wherein the position markers have adifferent optical characteristic than a surface of the detection zonethat is exposed for optical inspection.
 6. The system of claim 5,wherein the position markers are configured to encode positions alongthe assay test strip in a direction parallel to the flow path for thefluid sample.
 7. A diagnostic test system, comprising: an assay teststrip comprising a flow path for a fluid sample; a sample receiving zonecoupled to the flow path; a test label that specifically binds a targetanalyte; a control label that is free of any specific binding affinityfor the target analyte and has a different optical characteristic thanthe test label; a detection zone coupled to the flow path downstream ofthe sample receiving zone, the detection zone comprising a captureregion; an immobilized test reagent that specifically binds the targetanalyte in the capture region; and an immobilized control reagent thatspecifically binds the control label in the capture region, wherein theimmobilized test reagent and the immobilized control reagent areintermixed in the capture region; and a set of regularly spaced positionmarkers along an edge of the assay test strip distinct from the captureregion, the position markers having a different optical characteristicthan the surface of the detection zone; a reader comprising anillumination system that focuses a beam of light onto an area of thedetection zone, the cross-section of the focused beam of light on thesurface of the detection zone having at least one dimension at mostequal to the smallest of a first and a second dimension of the captureregion; and a data analyzer programmed to store light intensitymeasurements obtained from the position markers and to identify aposition of the capture region on the assay test strip using the storedlight intensity measurements and predetermined information correlatinglocations of regions of interest with a light intensity contrast patternproduced by the position markers.
 8. The system of claim 7, wherein theposition markers have a different reflectivity or fluorescence than thesurface of the detection zone.
 9. The system of claim 7, wherein thedata analyzer is programmed to identify ones of the light intensitymeasurements obtained from the capture region based on light intensitymeasurements obtained from at least one position marker on the teststrip.
 10. The system of claim 7, wherein the capture region has aplano-elliptical shape, and the illumination system is operable to focusa beam of light onto the plano-elliptical area of the capture region.11. The system of claim 7, wherein the test label fluoresces at a firstcharacteristic wavelength and the control label fluoresces at a secondcharacteristic wavelength different from the first characteristicwavelength.
 12. The system of claim 7, wherein the test label comprisesquantum dots of a first type and the control label comprises quantumdots of a second type.
 13. The system of claim 5, further comprising adata analyzer programmed to use the regularly spaced position markers todetermine when an illuminated area of the detection zone coincides withthe capture region.
 14. The system of claim 1, wherein the illuminationsystem focuses the beam of light in a shape and size configured toseparate the optical characteristic of the capture region from the restof the detection zone.
 15. The system of claim 1, wherein theillumination system scans the beam of light along a predetermined pathacross the detection zone, wherein the path includes a directiontransverse to the flow path for the fluid sample or a direction parallelto the flow path of the fluid sample.
 16. The system of claim 5, whereinthe regularly spaced position markers are configured to produce a lightintensity contrast pattern comprising a plurality of light intensitymeasurements, each light intensity measurement corresponding to theposition along the edge of the assay test strip associated with eachregularly spaced position marker and useable to identify a position ofthe capture region.
 17. The system of claim 7, wherein the data analyzeris programmed to use the regularly spaced position markers to determinewhen an illuminated area of the detection zone coincides with thecapture region.
 18. The system of claim 7, wherein the position markersare configured to encode positions along the assay test strip in adirection parallel to the flow path for the fluid sample.
 19. The systemof claim 1, wherein the test reagent and the control reagent areimmobilized in a combined test-and-control region of the detection zone.20. The system of claim 1, wherein the test reagent and the controlreagent are intermixed and immobilized in the capture region before thefluid sample is added to the flow path.
 21. The system of claim 7,wherein the test reagent and the control reagent are intermixed andimmobilized in the capture region before the fluid sample is added tothe flow path.